Sensitisation to House Dust Mite Component Der p 23 Is Associated with Severe Symptoms and Asthma in Allergic Rhinitis Patients.

INTRODUCTION: House dust mite (HDM) is an important source of airborne allergens in China as it contains several allergenic components that can cause allergic rhinitis (AR) and other allergic diseases. This study aimed to determine the clinical characteristics and disease severity in AR patients sensitised to different allergenic HDM components. METHODS: This was a retrospective study, which examined 129 patients who were first diagnosed with only HDM-induced AR at the Department of Allergy of Beijing Tongren Hospital from December 2019 to April 2021. Clinical characteristics and disease severity of the patients were assessed based on the sensitisation to specific Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) allergenic components, including Der p 1, Der p 2, Der p 23, Der f 1, and Der f 2, employing multiple correspondence analysis (MCA) with correspondence analysis chart of MCA. RESULTS: Among HDM-induced AR cases, the positive rate of Der p 1 was the highest (87.6%), followed by Der p 2 (78.3%), Der f 2 (76.64%), Der f 1 (68.2%), and Der p 23 (37.2%). Multiple correspondence analyses showed that sensitisation to Der p 23 was associated with severe AR symptoms and asthma; sensitisation to Der p 2, Der f 1, and Der f 2 was associated with moderate AR; and no sensitisation to Der p 23 was associated with mild AR. CONCLUSION: Der p 23 sensitisation is prevalent in northern China and may be associated with severe symptoms and asthma in AR patients.

Upregulation of Basonuclin1 Is Associated with p63-Involved Epithelial Barrier Impairment and Type-2 Helper T-cell Inflammation in Chronic Rhinosinusitis with Nasal Polyps.

BACKGROUND: Tumor protein p63 has been shown to be important for epithelial dysfunction, including epithelial barrier defects and mucosal inflammation, in the development of chronic rhinosinusitis with nasal polyps (CRSwNP). Basonuclin1 (BNC1), an epithelial-specific transcriptional factor, is a direct downstream target of p63 and thus might be involved in the pathogenesis of CRSwNP. OBJECTIVE: We sought to investigate whether BNC1 was associated with p63-mediated epithelial barrier defects and nasal mucosal inflammation in CRSwNP. METHODS: Nasal tissue biopsies were obtained from 91 patients to CRSwNP, 49 chronic rhinosinusitis without nasal polyps (CRSsNP) patients, and 28 control subjects. Immunohistochemistry and immunofluorescence staining were used to determine the distribution of BNC1 in tissues and localization in cells, respectively. Quantitative PCR was performed to detect the expression levels of BNC1, TP63, epithelial barrier proteins, and type-2 helper T-cell inflammation-related genes. RESULTS: BNC1 mRNA expression was significantly elevated in the tissues in CRSwNP patients compared with CRSsNP (1.96-fold, p = 0.0003) and control groups (2.40-fold, p < 0.0001). BNC1 staining was strongly positive in the nasal epithelium and co-localized with p63-positive epithelial cells. The expression of BNC1 mRNA was strongly correlated with TP63 mRNA level both in tissue biopsies (r = 0.78, p < 0.0001) and epithelial scrapings (r = 0.97, p < 0.0001). BNC1 expression was also positively correlated with epithelial barrier protein genes (CDH1, CLDN1, CLDN4, TJP1, and TJP2) and epithelial genes involved in TH2 inflammation (IL33, CCL26, CLC, and ALOX15). CONCLUSIONS: Overexpression of BNC1 may be associated with increased expression of TP63, and possibly contribute to the epithelial barrier defects and TH2 inflammation in CRSwNP.

Hypomethylation of the IL8 promoter in nasal epithelial cells of patients with chronic rhinosinusitis with nasal polyps.

BACKGROUND: IL-8 is an important chemokine implicated in the pathogenesis of chronic rhinosinusitis (CRS), but little is known about epigenetic regulation of IL8 in the pathogenesis of CRS. OBJECTIVE: We sought to investigate the relationship between the DNA methylation level in the IL8 proximal promoter and CRS in Han Chinese subjects. METHODS: Patients with chronic rhinosinusitis with nasal polyps (CRSwNP; n = 187), patients with chronic rhinosinusitis without nasal polyps (CRSsNP; n = 89), and control subjects (n = 57) were enrolled in 2 independent cohorts. Purified human nasal epithelial cells from each participant were assessed for percentage DNA methylation of CpG sites in the IL8 proximal promoter by using bisulfite pyrosequencing and for functional aspects of methylation status by using in vitro assays. RESULTS: DNA methylation of CpG sites 1, 2, and 3, respectively, in the IL8 proximal promoter was significantly decreased in human nasal epithelial cells of patients with CRSwNP compared with that in patients with CRSsNP (P < .001) and control subjects (P < .001). Percentage of DNA methylation of the CpG3 site was correlated negatively with both tissue eosinophilic cationic protein (P < .01) and myeloperoxidase (P < .05) levels. IL-1ß (P < .001) and TNF-α (P < .01) significantly increased IL8 expression accompanied by a reduction in methylation at the CpG3 site (P < .001). Electrophoretic mobility shift assays demonstrated that methylation status of CpG3 changed the binding of octamer-binding transcription factor 1 and nuclear factor κB. CONCLUSION: Decreased DNA methylation of particularly CpG sites in the IL8 proximal promoter might play a role in the pathogenesis of CRSwNP.

Blood and local eosinophil levels in chronic rhinitis: Observations during seasonal allergen exposure and non-exposure.

Background: Allergic rhinitis (AR) is a typical type 2 inflammatory disease and eosinophils play a critical role of cardinal effectors in type 2 inflammation. However, the distribution of eosinophils in patients with different subtypes of rhinitis and the effect of allergen exposure on them remain understudied. The aim of this study was to investigate the characteristics and factors influencing the distribution of systemic and local eosinophils in patients with non-AR (NAR), perennial AR (PAR), and seasonal AR (SAR), as well as the effect of seasonal allergen exposure levels on eosinophils. Methods: This was a population-based, cross-sectional observational study of consecutive chronic rhinitis (CR) outpatients who volunteered to participate in the survey during the spring pollen season and non-pollen season of 2023 in Beijing. All participants underwent serum allergen testing, blood routine examination, and nasal secretion smear cytology, and completed questionnaires mainly involving basic information, history review, and symptom assessment. Spring pollen dispersal concentration were considered. Results: A total of 558 CR patients eligible for enrollment were collected, including 198 NAR, 204 PAR, and 156 SAR patients. PAR had the highest blood eosinophil levels and the most severe overall nasal and ocular symptoms of SAR. Compared with subjects with blood eosinophil counts <0.3 × 109/L, those with ≥0.3 × 109/L had significantly more severe nasal and ocular symptoms and a significantly higher rate of comorbid asthma and allergic conjunctivitis. Blood eosinophil levels were significantly higher in SAR patients during the pollen season compared to the non-pollen season, and pollen concentrations were positively correlated with systemic and local eosinophil levels. Conclusions: Blood eosinophil levels varied in patients with different subtypes of rhinitis. Patients with high blood eosinophil levels had more severe overall nasal and ocular symptoms, and that blood eosinophils levels were significantly higher in patients with asthma or allergic conjunctivitis than patients without comorbidities. There was a positive trend between allergen exposure and systemic and local eosinophil levels. Further longitudinal cohort studies are still needed to better analyze the influence of eosinophil levels on the clinical traits of AR.

Long-term efficacy of HDM-SCIT in pediatric and adult patients with allergic rhinitis.

BACKGROUND: Subcutaneous immunotherapy (SCIT) is a well-validated and effective disease modification treatment for house dust mites (HDM)-induced allergic rhinitis (AR). Long-term post-treatment comparisons in children and adults treated with SCIT have rarely been published. This study aimed to evaluate the long-term efficacy of HDM-SCIT administered under a cluster schedule in children compared to adults. METHODS: This was an open-design, observational, long-term clinical follow-up study on children and adults with perennial AR treated with HDM-SCIT. The follow-up consisted of a three-year treatment duration plus a post-treatment follow-up of over three years. RESULTS: Patients in the pediatric (n = 58) and adult (n = 103) groups completed a post-SCIT follow-up of over three years. The total nasal symptom score (TNSS), combined symptom medication score (CSMS), and rhinoconjunctivitis quality-of-life questionnaire (RQLQ) score decreased significantly at T1 (three-year SCIT completed) and T2 (follow-up completed) in the pediatric and adult groups. In both groups, the improvement rate of TNSS (T0-T1) was moderately correlated with the baseline TNSS (r = 0.681, p < 0.001 and r = 0.477, p < 0.001 for children and adults, respectively). Only in the pediatric group, TNSS was significantly lower at T2 compared with that right after SCIT cessation (T1) (p = 0.030). CONCLUSIONS: Children and adults with HDM-induced perennial AR could achieve a sustainable post-treatment efficacy for over three years (up to 13 years) following a three-year SCIT. Patients with relatively severe nasal symptoms at baseline may benefit more from SCIT. Children who have completed an adequate course of SCIT may gain further improvement in nasal symptoms after SCIT cessation.

Establishing a novel 3D printing bioinks system with recombinant human collagen.

Bioinks are one of the key elements in realizing three-dimensional (3D) bioprinting. However, bioinks prepared from conventional collagen are hindered to their further applications due to concerns of collagen purity, unstable mechanical properties, and low solubility under neutralized conditions. This study aimed to develop a reliable UV-curable bioink system from a novel water-soluble recombinant human collagen (RHC). RHC was modified by methacrylic anhydride (MAA) and later crosslinked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) to obtain Pro-RHCMA. 1H nuclear magnetic resonance (1H NMR) confirmed the methacryloyl grafts, Fourier transform-infrared spectroscopy (FT-IR) illustrated the chemical crosslinking in producing the Pro-RHCMA. Internal morphology, mechanical properties and degradation of UV cured boinks were MAA and EDC/NHS modification-dependent. Photorheological properties and printability of the bioinks were determined. Cellular bioactivities were sustained within the printed bioinks, validating the bioinks biocompatibility in vitro. Finally, qRT-PCR revealed that the Pro-RHCMA bioinks provided a cell-friendly microenvironment for human umbilical vein endothelial cells (HUVECs) and human foreskin fibroblasts (HFFs), by supporting the expression of extracellular matrix (ECM) and angiogenesis-associated proteins, respectively. Taken together, this novel RHC-based bioink system shows great potential in tissue engineering and regenerative medicine.

Omalizumab is effective in the preseasonal treatment of seasonal allergic rhinitis.

BACKGROUND: To date no study has evaluated the efficacy of preseasonal omalizumab therapy with cost effective dose and at appropriate time point compared with standard medication in seasonal allergic rhinitis (SAR) patients. METHODS: This was a prospective randomized controlled open-label single-centre trial. 32 SAR patients were randomized to receive a single injection of omalizumab 300-mg approximately two weeks before start of the pollen period (PP) or medication therapy. All patients completed daily questionnaires; recording symptoms, medication use and quality of life (QoL) throughout the observation period. The primary efficacy parameter was the mean daily Combined Symptom and Medication Score (CSMS). RESULTS: Preseasonal omalizumab significantly reduced the changes of mean daily CSMS of nose during the PP (p < 0.001), peak pollen period (PPP) and PP after PPP (PPP-PP) (p = 0.002) and Post-PP (p = 0.009) compared to standard medication. The proportion of allergy symptoms-relieving medication-free days during PPP-PP was also significantly higher in preseasonal omalizumab-treated group (76.2(16.7-98.8))% than in medication-treated group (19.0(0-71.4))% (p = 0.030). Omalizumab could achieve the same nasal symptom control during the entire pollen season and better eye symptoms relieving results in PP (p = 0.046) and PPP-PP (p = 0.004) than medication treatment. Significantly greater improvement in QoL was also obtained with omalizumab-pretreatment during the PP (p = 0.037) and PPP-PP (p = 0.004). CONCLUSIONS: Administration of a single injection of 300 mg omalizumab two weeks before start of the pollen season achieves better overall control of symptoms and QoL, with significantly reduced allergy symptoms-relieving medication usage, compared with standard pharmacotherapy in SAR patients.

Construction of a BMP-2 gene delivery system for polyetheretherketone bone implant material and its effect on bone formation in vitro.

Polyetheretherketone (PEEK) has been widely investigated for improving its biological inert to enable it to achieve stronger osteogenic capability and to be a promising material in implant fields. The most important mechanism that makes a successful implantation is osteointegration. Surface modification is an appropriate method to maintain the excellent mechanical properties of PEEK and simultaneously endow PEEK certain biological characters. In this work, we attempted to shape the nano-topography of PEEK surface by nitrogen low-temperature plasma and polydopamine coating on the surface as a secondary reaction platform to bond the aminated poly (lactic-co-glycolic acid) (PLGA) microspheres encapsulating the BMP-2 gene for enhancing the biological activity. Scanning electron microscope, atomic force microscopy, X-ray photoelectron spectroscopy and water contact angle (CA) measurements were applied to characterize the surface of modified or untreated PEEK. Surface characterization showed that the modification was successfully performed on PEEK including a rougher and more hydrophilic surface with nanotopographic features. The influence on cell adhesion, proliferation and differentiation was evaluated by culturing of rat bone marrow mesenchymal stem cells on different modified PEEK substrates in vitro. The biological results indicated that the low-temperature plasma treatment and PDA-coating on PEEK significantly promoted cell adhesion and proliferation. And the osteogenic differentiation was effectively improved by BMP-2 gene releasing from PLGA-NH2 microspheres. The results showed that this novel biological surface modification endowed PEEK with outstanding bioactivity and osteogenic ability, providing a theoretical basis for application in the field of implantation.

Construction and performance evaluation of a sustained release implant material polyetheretherketone with antibacterial properties.

OBJECTIVE: This study aimed to construct a tightly binding antibiotic sustained release system on the polyetheretherketone (PEEK) surface and investigate the cellular activity and antibacterial properties of the new oral implant materials. METHODS: Low-temperature argon plasma under certain parameters was used to prepare P-PEEK with nano-topology, and chemical deposition technology was adopted to form a polydopamine (PDA) coating on the PEEK surface to build a biological binding platform, PDA/P-PEEK. Subsequently, vancomycin gelatin nanoparticles (Van-GNPs) were prepared by two-step desolvation method. Finally, Van-GNPs were combined with PEEK implant material surface to form a new composite material, Van-GNPs/PEEK. scanning electron microscope (SEM), atomic force microscope (AFM), energy dispersive spectrometer (EDS), and contact angle tester were used to comprehensively characterize the materials. The in vitro release test of Van was performed by dynamic dialysis with ultraviolet spectrophotometer. The cell cytotoxicity and adhesion tests were studied by mouse embryonic osteoblasts. The antibacterial properties were evaluated by bacterial adhesion test, plate colony counting, and antimicrobial ring test with Staphylococcus aureus and Streptococcus mutans. RESULTS: PEEK was treated with low-temperature argon plasma and attached to PDA to form a biological binding platform. The synthesized Van-GNPs were smooth, round, with uniform particle size distribution, and bound to PEEK to form a new composite material, which can release Van constantly. Cell experiments showed that Van-GNPs/PEEK had no cytotoxicity and had good interaction with osteoblasts. Bacterial experiments showed that surface conjugation with Van-GNPs could significantly improve the antibacterial performance of PEEK against S. aureus and S. mutans. SIGNIFICANCE: This study demonstrated that Van-GNPs/PEEK have good cellular compatibility and autonomous antibacterial properties, which provide a theoretical basis for the wide application of PEEK in the field of stomatology.

Replication study of susceptibility variants associated with allergic rhinitis and allergy in Han Chinese.

BACKGROUND: Allergic rhinitis (AR) is believed to be a complex genetic disease. The last decade has been marked by the publication of more than 20 genome-wide association studies (GWASs) of AR and associated allergic phenotypes and allergic diseases, which have shown allergic diseases and traits to share a large number of genetic susceptibility loci. The aim of present study was therefore to investigate the highly replicated allergy related genes and variants as candidates for AR in Han Chinese subjects. METHODS: A total of 762 AR patients and 760 control subjects were recruited, and a total of 58 susceptible variants previously reported to be associated with allergic traits were choose for replication. RESULTS: Logistic regression analyses revealed that in the co-dominant-effect model as assessed by the AIC, compared with wild-type carriers, significant AR risk were associated with rs9865818 in LPP (P = 0.029, OR = 1.469 for GG vs. AA); rs6554809 in DNAH5 (P = 0.000, OR = 1.597 for TC vs. CC); rs1438673 in WDR36-CAMK4 loci (P = 0.037, OR = 1.396 for CC vs.TT), rs7775228 in HLA region (P = 0.000, OR = 1.589 for TC vs.TT), rs7203459 in CLEC16A (P = 0.025, OR = 0.731 for TC vs. TT). CONCLUSION: We replicated Han Chinese AR-specific susceptibility loci in LPP, DNAH5, HLA, CLEC16A and WDR36-CAMK4. Further understanding the molecular mechanisms underlying these associations may provide new insights into the etiology of allergic disease.

Association between methylation in nasal epithelial TSLP gene and chronic rhinosinusitis with nasal polyps.

BACKGROUND: This study was performed to determine whether there was any association between abnormal DNA methylation of a thymic stromal lymphopoietin (TSLP) locus and pathogenesis of chronic rhinosinusitis (CRS). METHODS: A total of 48 CRS patients with nasal polyps (CRSwNP), 28 CRS patients without nasal polyps (CRSsNP) and 21 control subjects were enrolled into the study; and evaluated for serum total IgE level, olfactory score and nasal resistance. Samples were obtained from nasal polyps of CRSwNP patients, ethmoid mucosae of CRSsNP patients and inferior turbinate (IT) mucosa of control subjects during surgery, and used to isolate purified primary human nasal epithelial cells (HNECs). Genomic DNA was extracted from purified primary HNECs of each subject and DNA methylation ratios for a selected region of the TSLP gene were screened the using MassARRAY EpiTYPER. RESULTS: A total of 17 CpG units were analyzed; of which two CpG units (CpG3 and 22:23:24) had increased methylation ratios in the CRSwNP patients compared to the CRSsNP and control subjects after correction for false discovery rate (FDR) (Q < 0.1). The methylation ratios at both CpG3 and CpG22:23:24 units were positively correlated with olfactory score (r = 0.41, P = 0.0001; r = 0.25, P = 0.021) and unilateral nasal resistance at 75 Pa (r = 0.24, P = 0.04; r = 0.24, P = 0.036) and 150 Pa (r = 0.34, P = 0.004; r = 0.25, P = 0.031). Total nasal resistance at 75 Pa/150 Pa or serum total IgE levels were not correlated with the methylation ratios at either CpG unit. CONCLUSIONS: Increased DNA methylation at the TSLP locus is likely to be associated with CRSwNP pathogenesis; however these findings need to be confirmed in larger multicentre group studies.